A reproducible microcosm biofilm model of subgingival microbial communities

Mostajo, Mercedes Fernandez y; Exterkate, R.A.M.; Buijs, Mark J.; Beertsen, W.; Weijden, G.A. van der; Zaura, Egija; Crielaard, Wim

doi:10.1111/jre.12473

Cited by 25 publications

(25 citation statements)

References 62 publications

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“…Recently, we could show that microcosm biofilms resembling subgingival periodontitis-associated biofilms can be cultured in a model using subgingival plaque sampled from periodontitis patients as an inoculum. This model supported the growth of fastidious anaerobic and proteolytic Gram-negative bacteria [25]. However, for practical reasons, a limited amount of subgingival plaque is available from a single patient.…”

Section: Introductionmentioning

confidence: 84%

“…Yet another option combining advantageous features from both models described above is to take intra-oral microbiological samples from patients, e.g. plaque or saliva, and use these as ex vivo inocula to grow so-called microcosm biofilms in vitro [19,[23][24][25][26][27]. These microcosm biofilms inoculated from patient samples are closer to the complex in vivo situation as compared to in vitro biofilms from defined consortia and exhibit easier handling and less dependence on participants as compared to biofilms grown in situ [19,24].…”

Section: Introductionmentioning

confidence: 99%

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Microcosm biofilms cultured from different oral niches in periodontitis patients

Zaura

Brandt

Buijs

et al. 2018

Journal of Oral Microbiology

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Objective: Periodontal diseases are triggered by dysbiotic microbial biofilms. Therefore, it is essential to develop appropriate biofilm models. Aim of the present study was to culture microcosm biofilms inoculated from different niches in periodontitis patients and compare their microbial composition to those inoculated from subgingival plaque.Methods: Saliva, subgingival plaque, tongue and tonsils were sampled in five periodontitis patients to serve as inocula for culturing biofilms in vitro in an active attachment model. Biofilms were grown for 14 or 28 d and analyzed for their microbial composition by 16S rDNA sequencing.Results: As classified by HOMD, all biofilms were dominated by periodontitis-associated taxa, irrespective which niche had been used for inoculation. There was a low similarity between 14 d biofilms and their respective inocula (Bray-Curtis similarity 0.26), while biofilms cultured for 14 and 28 d shared high similarity (0.69). Principal components analysis showed much stronger clustering per patient than per niche indicating that the choice of patients may be more crucial than choice of the respective niches in these patients.Conclusion: Saliva, tongue scrapings or tonsil swabs may represent sufficient alternative inocula for growing microcosm biofilms resembling periodontitis-associated microbial communities in cases when sampling subgingival plaque is not possible.

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Section: Introductionmentioning

confidence: 84%

Section: Introductionmentioning

confidence: 99%

Microcosm biofilms cultured from different oral niches in periodontitis patients

Zaura

Brandt

Buijs

et al. 2018

Journal of Oral Microbiology

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“…The AAA model offers the advantage of active attachment (rather than sedimentation) of the bacteria onto the substrate, and also facilitates controlling the periods which the biofilms are exposed to the tested compounds [34]. Choice of inoculum and growth conditions are crucial aspects for microcosm biofilms [40,[42][43][44][45]. In the present study, human saliva was chosen as inoculum source because it can be collected easier and in higher quantities as compared with dental plaque [40].…”

Section: Discussionmentioning

confidence: 99%

Limited antimicrobial efficacy of oral care antiseptics in microcosm biofilms and phenotypic adaptation of bacteria upon repeated exposure

et al. 2020

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Objectives The aims of this study were to investigate the antimicrobial efficacy of antiseptics in saliva-derived microcosm biofilms, and to examine phenotypic adaption of bacteria upon repeated exposure to sub-inhibitory antiseptic concentrations. Methods Saliva-derived biofilms were formed mimicking caries- or gingivitis-associated conditions, respectively. Microbial compositions were analyzed by semiconductor-based 16S rRNA sequencing. Biofilms were treated with CHX, CPC, BAC, ALX, and DQC for 1 or 10 min, and colony forming units (CFU) were evaluated. Phenotypic adaptation of six selected bacterial reference strains toward CHX, CPC, and BAC was assessed by measuring minimum inhibitory concentrations (MICs) over 10 passages of sub-inhibitory exposure. Protein expression profiles were investigated by SDS-PAGE. Results Both biofilms showed outgrowth of streptococci and Veillonella spp., while gingivitis biofilms also showed increased relative abundances of Actinomyces, Granulicatella, and Gemella spp. Antiseptic treatment for 1 min led to no relevant CFU-reductions despite for CPC. When treated for 10 min, CPC was most effective followed by BAC, ALX, CHX, and DQC. Stable adaptations with up to fourfold MIC increases were found in E. coli toward all tested antiseptics, in E. faecalis toward CHX and BAC, and in S. aureus toward CPC. Adapted E. coli strains showed different protein expression as compared with the wildtype strain. Conclusion Antiseptics showed limited antimicrobial efficacy toward mature biofilms when applied for clinically relevant treatment periods. Bacteria showed phenotypic adaptation upon repeated sub-inhibitory exposure. Clinical relevance Clinicians should be aware that wide-spread use of antiseptics may pose the risk of inducing resistances in oral bacteria.

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“…We do note that our inoculum was distinct from general descriptions of periodontitis communities. Fusobacterium is a common genus found in both health and disease communities (Colombo & Tanner, 2019) and its relative abundance can be over 30% of a periodontitis sample (Baraniya et al., 2020; Fernandez y Mostajo et al., 2017). However, just one species of Fusobacterium was present in the current inoculum, making up only 0.07% of the community.…”

Section: Resultsmentioning

confidence: 99%

Modified SHI medium supports growth of a disease‐state subgingival polymicrobial community in vitro

Lamont

Gadkari

Kerns

et al. 2020

Molecular Oral Microbiology

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Developing a laboratory model of oral polymicrobial communities is essential for in vitro studies of the transition from healthy to diseased oral plaque. SHI medium is an enriched growth medium capable of supporting in vitro biofilms with similar diversity to healthy supragingival inocula; however, this medium does not maintain the diversity of gram‐negative bacteria more associated with subgingival plaque. Here, we systematically modified SHI medium components to investigate the impacts of varying nutrients and develop a medium capable of supporting a specific disease‐state subgingival community. A diseased subgingival plaque sample was inoculated in SHI medium with increasing concentrations of sucrose (0%, 0.1%, 0.5%), fetal bovine serum (FBS) (0%, 10%, 20%, 30%, 50%), and mucin (0.1, 2.5, 8.0 g/L) and grown for 48 hrs, then the 16S rRNA profiles of the resulting biofilms were examined. In total, these conditions were able to capture 89 of the 119 species and 43 of the 51 genera found in the subgingival inoculum. Interestingly, biofilms grown in high sucrose media, although dominated by acidogenic Firmicutes with a low final pH, contained several uncultured taxa from the genus Treponema, information that may aid culturing these periodontitis‐associated fastidious organisms. Biofilms grown in a modified medium (here named subSHI‐v1 medium) with 0.1% sucrose and 10% FBS had a high diversity closest to the inoculum and maintained greater proportions of many gram‐negative species of interest from the subgingival periodontal pocket (including members of the genera Prevotella and Treponema, and the Candidate Phyla Radiation phylum Saccharibacteria), and therefore best represented the disease community.

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A reproducible microcosm biofilm model of subgingival microbial communities

Abstract: This biofilm model allows reproducible production of complex subgingival microbial communities.

Cited by 25 publications

References 62 publications

Microcosm biofilms cultured from different oral niches in periodontitis patients

Microcosm biofilms cultured from different oral niches in periodontitis patients

Limited antimicrobial efficacy of oral care antiseptics in microcosm biofilms and phenotypic adaptation of bacteria upon repeated exposure

Modified SHI medium supports growth of a disease‐state subgingival polymicrobial community in vitro

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